Download InviMag Blood DNA Mini Kit/ KFmL User manual

User manual
InviMag® Blood DNA Mini Kit/ KFmL
for use on KingFisher®mL, Thermo Fisher Scientific
for automated purification of total DNA from up to 200 µl of whole blood samples, buffy
coat, non mammalian blood, bone marrow, and swabs with magnetic beads
IVD
REF 2431110x00
STRATEC Molecular GmbH, D-13125 Berlin
Instruction InviMag® Blood DNA Mini Kit/ KFmL
The InviMag® Blood DNA Mini Kit/ KFmL combines the advantages of the innovative Invisorb®
technology with easy handling of magnetic particles for a very efficient and reliable isolation of
nucleic acids with a high purity.
The DNA-binding magnetic particles are characterized by a high surface area, a uniform size
distribution, and good suspension stability. They are highly suitable for high-throughput
processing.
The InviMag® Blood DNA Mini Kit/ KFmL for isolation and purification of total (genomic and
mitochondrial) DNA from whole blood samples, buffy coat, non-mammalian blood, cerebrospinal
fluid (CSF), bone marrow, and swabs in a single well format for up to 15 samples per run is
designed for an optimal use on the KingFisher® mL workstation from Thermo Scientific. The
interplay of the DNA extraction and purification chemistry provided by the InviMag® Blood DNA
Mini Kit/ KFmL with the KingFisher machine was intensely tested and validated.
The kit is neither suitable for isolation of DNA from stool samples, tissue samples, bacteria,
fungi, plants or viruses nor for purification of RNA.
IVD
Compliance with EU Directive 98/79/EC on in vitro medical devices.
Not for in vitro diagnostic use in countries where the EU Directive 98/79/EC on in vitro medical devices is not
recognized.
Trademarks: InviMag®, Invisorb®. Registered marks, trademarks, etc. used in this document, even when not specifically marked as
such, are not to be considered unprotected by law.
The Invisorb® technology is covered by patents and patent applications: US 6,110363, US 6,043,354, US 6,037,465, EP 0880535,
WO 9728171, WO 9534569, EP 0765335, DE 19506887, DE 10041825.2, WO 0034463.
InviMag® and Invisorb® are registered trademarks of STRATEC Biomedical AG.
The PCR process is covered by US Patents 4,683,195, and 4,683,202 and foreign equivalents owned by Hoffmann-La Roche AG.
© 2015 STRATEC Molecular, all rights reserved.
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InviMag Blood DNA Mini Kit/ KFmL 0515
Content
Kit contents of InviMag® Blood DNA Mini Kit/ KFmL
3
Kit contents of InviMag® Blood DNA Mini Kit/ KFmL / w/o plastic
4
Symbols
5
Storage
5
Quality control
5
Intended use
6
Product use limitation
6
Safety information
7
Product characteristics of the InviMag® Blood DNA Mini Kit/ KFmL
8
Sampling and sample storage of the starting material
9
Principle and procedure
9
Yield and quality of genomic DNA
10
Reagents and equipment to be supplied by user
10
Important points before starting a protocol
11
Preparing reagents and buffers
11
®
Scheme of the InviMag Blood DNA Mini Kit/ KFmL
12
Protocol 1: Isolation of genomic DNA from up to 200 µl of whole blood or 1–30 µl buffy coat
13
Protocol 2: Isolation of genomic DNA from up to 25 µl of non mammalian blood
13
Protocol 3: Isolation of genomic DNA from CSF and bone marrow
14
Protocol 4: Isolation of genomic DNA from swabs or rinsed liquid from swabs
14
Starting a run - Preliminary Steps to process the sample onto the KingFisher System
16
For self programming the KFmL system
17
Troubleshooting
19
Appendix
20
General notes on handling DNA
21
Ordering information
22
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InviMag Blood DNA Mini Kit/ KFmL 0515
Kit contents of InviMag® Blood DNA Mini Kit/ KFmL
Store the SNAP Solution at 4°C! Store dissolved Proteinase K at -20°C!
Store all other kit components at room temperature (RT)!
15 extractions
75 extractions
300extractions
Catalogue No.
2431110100
2431110200
2431110400
Lysis Buffer A
5 ml
20 ml
70 ml
for 0.5 ml
working solution
for 2 ml
working solution
for 7 ml
working solution
0.5 ml
2 x 1 ml
7 ml
3 ml
(final volume 10 ml)
2 x 9 ml
(final volume 2 x 30 ml)
2 x 24 ml
(final volume 2 x 80 ml)
Proteinase K
(working solution)
SNAP Solution
Binding Buffer B6
Wash Buffer I
7.5 ml
2 x 30 ml
2 x 80 ml
(final volume 15 ml)
(final volume 2 x 60 ml)
(final volume 2 x 160 ml)
18 ml
45 ml
3 x 60 ml
(final volume 60 ml)
(final volume 150 ml)
(final volume 3 x 200 ml)
2 x 2 ml
20 ml
70 ml
KingFisher mL Tip Combs
3
15
60
KingFisher mL Tube Strips
15
5 x 15
300
1.5 ml Receiver Tubes
15
5 x 15
6 x 50
Manual
1
1
1
Add 7 ml 99.7%
Isopropanol to the Binding
Buffer B6. Mix by intensive
shaking by inverting for 1
min. Shortly before use mix
by inverting several times.
Add 21 ml 99.7%
Isopropanol to each
Binding Buffer B6. Mix by
intensive shaking by
inverting for 1 min. Shortly
before use mix by inverting
several times.
Add 56 ml 99.7%
Isopropanol to each
Binding Buffer B6. Mix by
intensive shaking by
inverting for 1 min. Shortly
before use mix by inverting
several times.
Dilute Proteinase K by
addition of 2 ml ddH2O, mix
thoroughly until completely
dissolving and store at 20°C!
Dilute Proteinase K by
addition of 7 ml ddH2O, mix
thoroughly until completely
dissolving and store at 20°C!
Add 30 ml of 96-100%
ethanol to the bottle Wash
Buffer I, mix thoroughly and
always keep the bottle firmly
closed!
Add 80 ml of 96-100%
ethanol to each bottle Wash
Buffer I, mix thoroughly and
always keep the bottle firmly
closed!
Add 105 ml of 96-100%
ethanol to the bottle Wash
Buffer II, mix thoroughly and
always keep the bottle firmly
closed!
Add 140 ml of 96-100%
ethanol to each bottle Wash
Buffer II, mix thoroughly and
always keep the bottle firmly
closed!
Wash Buffer II
Elution Buffer
Initial steps
Dilute Proteinase K by
addition of 0.5 ml ddH2O,
mix thoroughly until
completely dissolving and
store at -20°C!
Add 7.5 ml of 96-100%
ethanol to the bottle Wash
Buffer I, mix thoroughly and
always keep the bottle firmly
closed!
Add 42 ml of 96-100%
ethanol to the bottle Wash
Buffer II, mix thoroughly and
always keep the bottle firmly
closed!
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InviMag Blood DNA Mini Kit/ KFmL 0515
Kit contents of InviMag® Blood DNA Mini Kit/ KFmL/ w/o plastic
Store the SNAP Solution at 4°C! Store dissolved Proteinase K at -20°C!
Store all other kit components at room temperature (RT)!
15 extractions
75 extractions
300extractions
Catalogue No.
2431110150
2431110250
2431110450
Lysis Buffer A
5 ml
20 ml
70 ml
for 0.5 ml
working solution
for 2 ml
working solution
for 7 ml
working solution
0.5 ml
2 x 1 ml
7 ml
3 ml
(final volume 10 ml)
2 x 9 ml
(final volume 2 x 30 ml)
2 x 24 ml
(final volume 2 x 80 ml)
Proteinase K
(working solution)
SNAP Solution
Binding Buffer B6
Wash Buffer I
7.5 ml
2 x 30 ml
2 x 80 ml
(final volume 15 ml)
(final volume 2 x 60 ml)
(final volume 2 x 160 ml)
18 ml
45 ml
3 x 60 ml
(final volume 60 ml)
(final volume 150 ml)
(final volume 3 x 200 ml)
2 x 2 ml
20 ml
70 ml
1.5 ml Receiver Tubes
15
5 x 15
6 x 50
Manual
1
1
1
Wash Buffer II
Elution Buffer
Initial steps
Add 7 ml 99.7%
Isopropanol to the Binding
Buffer B6. Mix by intensive
shaking by inverting for 1
min. Shortly before use mix
by inverting several times.
Dilute Proteinase K by
addition of 0.5 ml ddH2O,
mix thoroughly until
completely dissolving and
store at -20°C!
Add 7.5 ml of 96-100%
ethanol to the bottle Wash
Buffer I, mix thoroughly
and always keep the bottle
firmly closed!
Add 42 ml of 96-100%
ethanol to the bottle Wash
Buffer II, mix thoroughly and
always keep the bottle firmly
closed!
Add 21 ml 99.7%
Isopropanol to each
Binding Buffer B6. Mix by
intensive shaking by
inverting for 1 min. Shortly
before use mix by inverting
several times.
Add 56 ml 99.7%
Isopropanol to each
Binding Buffer B6. Mix by
intensive shaking by
inverting for 1 min. Shortly
before use mix by inverting
several times.
Dilute Proteinase K by
addition of 2 ml ddH2O, mix
thoroughly until completely
dissolving and store at 20°C!
Dilute Proteinase K by
addition of 7 ml ddH2O, mix
thoroughly until completely
dissolving and store at 20°C!
Add 30 ml of 96-100%
ethanol to the bottle Wash
Buffer I, mix thoroughly and
always keep the bottle firmly
closed!
Add 80 ml of 96-100%
ethanol to each bottle Wash
Buffer I, mix thoroughly and
always keep the bottle firmly
closed!
Add 105 ml of 96-100%
ethanol to the bottle Wash
Buffer II, mix thoroughly and
always keep the bottle firmly
closed!
Add 140 ml of 96-100%
ethanol to each bottle Wash
Buffer II, mix thoroughly and
always keep the bottle firmly
closed!
Plastic to be supplied by
user (see order information)
KingFisher mL Tip Combs
3
15
60
KingFisher mL Tube Strips
15
5 x 15
300
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InviMag Blood DNA Mini Kit/ KFmL 0515
Symbols
Manufacturer
Lot number
Catalogue number
Expiry date
Consult operating instructions
Temperature limitation
Do not reuse
Storage
All buffers and kit contents of the InviMag® Blood DNA Mini Kit/ KFmL except dissolved
Proteinase K and SNAP Solution should be stored at room temperature* and are stable for at
least 12 months under these conditions.
Proteinase K: Dissolved Proteinase K must be stored at -20°C. Dividing the Proteinase K into
aliquots and storage at -20°C is recommended.
SNAP Solution: SNAP Solution should be stored at 4°C.
Wash Buffers: Wash Buffers charged with ethanol should be stored at room temperature* and
should be appropriate sealed. If there are any precipitates visible within the provided solutions
solve them by carefully warming up to room temperature (up to 30°C).
Room temperature (RT) is defined as range from 15-30°C.
Quality control and product warranty
STRATEC Molecular warrants the correct function of the InviMag® Blood DNA Mini Kit/ KFmL
for applications as described in this manual. Purchaser must determine the suitability of the
product for its particular use. Should any product fail to perform the applications as described in
the manual, STRATEC Molecular will check the lot and if STRATEC Molecular investigates a
problem in the lot, STRATEC Molecular will replace the product free of charge.
STRATEC Molecular reserves the right to change, alter, or modify any product to enhance its
performance and design at any time.
In accordance with STRATEC Molecular’s ISO 9001-2000 and ISO EN 13485 certified Quality
Management System the performance of all components of the InviMag® Blood DNA Mini Kit/
KFmL have been tested separately against predetermined specifications routinely on lot-to-lot to
ensure consistent product quality.
If you have any questions or problems regarding any aspects of InviMag® Blood DNA Mini Kit/
KFmL or other STRATEC Molecular products, please do not hesitate to contact us. A copy of
STRATEC Molecular’s terms and conditions can be obtained upon request or are presented at
the STRATEC Molecular webpage.
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InviMag Blood DNA Mini Kit/ KFmL 0515
For technical support or further information please contact:
from Germany: +49-(0)30-9489-2901/ 2910
from abroad: +49-(0)30-9489-2907
or contact your local distributor.
Intended use
The InviMag® Blood DNA Mini Kit/ KFmL is designed for semi-automated extraction and
purification of total (genomic and mitochondrial) DNA from up to 15 whole blood or blood related
samples using magnetic beads and the KingFisher mL instrument. The nucleic acid isolation
protocol is suitable for routinely automated preparation of DNA from fresh or frozen whole blood
sample, buffy coat, non mammalian blood, cerebrospinal fluid (CSF), bone marrow, and swabs.
For reproducible and high yields an appropriate sample storage is essential (see “Sampling and
storage of the starting material”, page 9). Common blood collection tubes (not provided) and
anticoagulants (EDTA, citrate, but not heparin) can be used to assemble a set of blood
samples. All utilities (reagents and plastic ware) necessary for preparation of total DNA are
provided by the InviMag® Blood DNA Mini Kit/ KFmL in different package sizes.
The procedure of the InviMag® Blood DNA Mini Kit/ KFmL is optimized for the isolation of total
DNA from up to 200 µl starting material. For samples of a smaller volume than 200 µl please
adjust to a total sample volume of 200 µl using distilled water or 1x PBS before starting a
purification protocol.
THE PRODUCT IS INDENTED FOR USE BY PROFESSIONALS ONLY, SUCH AS
TECHNICIANS, PHYSICIANS AND BIOLOGISTS TRAINED IN MOLECULAR BIOLOGICAL
TECHNIQUES. It is designed to be used with any downstream application employing enzymatic
amplification or other enzymatic modifications of DNA followed by signal detection or
amplification. Any diagnostic results generated by using the sample preparation procedure in
conjunction with any downstream diagnostic assay should be interpreted with regard to other
clinical or laboratory findings.
To minimize irregularities in diagnostic results, adequate controls for downstream applications
should be used.
The kit is in compliance with EU Directive 98/79/EC on in vitro medical devices. But it is not for in vitro diagnostic use
in countries where the EU Directive 98/79/EC on in vitro medical devices is not recognized.
Product use limitation
The kit is neither validated for the isolation of DNA from stool samples, tissue, bacteria, fungi or
viruses, nor for isolation and purification of RNA.
The included chemicals are only useable once.
Differing of starting material or flow trace may lead to inoperability; therefore neither a warranty
nor guarantee in this case will be given, neither implied nor express.
The user is responsible to validate the performance of the STRATEC Molecular product for any
particular use. STRATEC Molecular does not provide for validation of performance characteristics
of the product with respect to specific applications. STRATEC Molecular products may be used
e.g.in clinical diagnostic laboratory systems conditioned upon the complete diagnostic system of
the laboratory the laboratory has been validated pursuant to CLIA’ 88 regulations in the U.S. or
equivalents in other countries.
All products sold by STRATEC Molecular are subject to extensive quality control procedures
(according to ISO 9001-2000 and ISO EN 13485) and are warranted to perform as described
herein. Any problems, incidents or defects shall be reported to STRATEC Molecular immediately
upon detection thereof.
The chemicals and the plastic parts are for laboratory use only; they must be stored in the
laboratory and must not be used for purposes other than intended.
The product with its contents is unfit for consumption.
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InviMag Blood DNA Mini Kit/ KFmL 0515
Safety information
When and while working with chemicals, always wear a suitable lab coat, disposable gloves, and
protective goggles!
Avoid skin contact! Adhere to the legal requirements for working with biological material!
For more information, please consult the appropriate material safety data sheets (MSDS). These
are available online in convenient and compact PDF format at www.stratec.com for each
STRATEC Molecular product and its components. If buffer bottles are damaged or leaking, WEAR
GLOVES, AND PROTECTIVE GOGGLES when discarding the bottles in order to avoid any
injuries.
STRATEC Molecular has not tested the liquid waste generated by the InviMag® Blood DNA
Mini Kit/ KFmL procedures for residual infectious materials. Contamination of the liquid waste
with residual infectious materials is highly unlikely, but cannot be excluded completely.
Therefore, liquid waste has be considered infectious and be handled and discarded according to
local safety regulations.
European Community risk and safety phrases for the components of the InviMag® Blood DNA
Mini Kit/ KFmL to which they apply, are listed below as follows:.
Wash Buffer I contains guanidine thiocyanate which is an irritant.
Lysis Buffer A
danger
H-319 P305-351-338
Proteinase K
Wash Buffer I
danger
H315-319-334-335 P280-305-351-338-310-405
warning
H302-312-332-412 EUH032 P273
H319:
H315:
H334:
H335:
H302:
H312:
H332:
H412:
EUH032:
P305+P351+P338:
P280:
P310:
P405:
P273:
Causes serious eye irritation.
Causes skin irritation.
May cause allergy or asthma symptoms or breathing difficulties if inhaled.
May cause respiratory irritation.
Harmful if swallowed.
Harmful in contact with skin.
Harmful if inhaled.
Harmful to aquatic life with long lasting effects.
Contact with acids liberates very toxic gas.
IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if
present and easy to do. Continue rinsing.
Wear protective gloves/protective clothing/eye protection/face protection.
Immediately call a POISON CENTER or doctor/physician.
Store locked up.
Avoid release to the environment.
Emergency medical information can be obtained 24 hours a day from infotrac:
outside of USA:
in USA :
1 – 352 – 323 – 3500
1 – 800 – 535 – 5053
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InviMag Blood DNA Mini Kit/ KFmL 0515
Product characteristics of the InviMag® Blood DNA Mini Kit/ KFmL
The InviMag® Blood DNA Mini Kit/ KFmL procedure the ideal tool for an efficient DNA
extraction and purification from fresh or frozen whole blood samples, non-mammalian blood,
buffy coat, CST, bone marrow, and swabs in a single well format for up to 15 samples per run
using magnetic beads and the KingFisher mL.
Starting material
Yield
Time
1-200 µl fresh, frozen or old human or
other mammalian whole blood;
1-200 µl cerebrospinal fluid;
1–30 µl buffy coat;
1–25 µl fresh, frozen or old non
mammalian blood;
1–20 µl bone marrow;
swabs;
up to 200 µl rinsed liquid from swab
up to 10 µg (in average abou 30 min
6 µg) depends on amount of (without lysis)
lymphocytes, sample source
sample transport, sample
storage, and age of the
sample
Ratio
A260 : A 280
1.7 – 2.0
The DNA isolation process is based on the interaction of nucleic acids with coated magnetic
particles under adapted buffer conditions. After lysis, the KingFisher mL performs all steps of the
DNA purification procedure automatically without any user intervention. The procedure requires
minimal interaction by the user, thus allowing safe handling of potentially infectious samples.
Sample cross-contaminations and reagent cross-over are effectively eliminated by this
automated purification process.
The KingFisher® instruments use magnetic rods to transport the DNA-binding magnetic particles
through the various purification phases: binding-washing-elution. The volume of buffers and
other liquids necessary for DNA isolation is reduced to a minimum.
After an external sample specific cell lysis using Lysis Buffer A and Proteinase K, optimal
binding conditions are adjusted upon addition of Binding Solution B6. The total DNA bound to
the simultaneously added magnetic particles is separated from solution by the magnetic rods
controlled by the KingFisher instrument. Subsequent to the three washing steps of the particle
bound nucleic acids, the DNA is finally eluted in Elution Buffer.
Due to the high purity, the eluted (genomic and mitochondrial) DNA is ready-to-use for a broad
panel of downstream applications:
○ PCR*
○ Restriction Enzyme Digestion
○ HLA Typing
○ Southern Blot.
The InviMag® Blood DNA Mini Kit/ KFmL is supplied with a comprehensive manual describing
four protocols (page 13-14) for DNA purification from different sources.
For the isolation of genomic DNA using magnetic particles in a 96 well format, STRATEC
Molecular offers the InviMag® Blood DNA Mini Kit/ KF96/ KFflex96 for use on a KF96 /
KFflex96 instrument. For the isolation of DNA from a single blood sample STRATEC Molecular
offers the Invisorb® Spin Blood Mini Kit or for 8-96 samples the Invisorb® DNA Blood Mini
HTS 96 Kits for use on a centrifuge, vacuum manifold or other robotic stations (see “Ordering
Information”, page 22).
For further information please contact: Phone +49 (0) 30 9489 2901, 2910 in Germany and
from foreign countries phone +49 (0) 30 9489 2907 or your local distributor.
* The PCR process is covered by US Patents 4,683,195, and 4,683,202 and foreign equivalents owned by HoffmannLa Roche AG.
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InviMag Blood DNA Mini Kit/ KFmL 0515
Sampling and sample storage of the starting material
For reproducible and high yields appropriate sample storage is essential. Yields may be varying
from sample to sample depending on factors such as health of the donor, sample age, kind of
sample, transport and storage conditions.
Blood and Buffy Coat:
Best results are obtained using fresh blood samples. Mammalian blood samples (stabilized with
EDTA or citrate but not heparin) can be stored at room temperature for up to 2-3 hours. For
short-term storage (up to 24 h) samples should be stored at 4°C. For long-term storage, we
recommend freezing samples at -20°C or -80°C. Multiple thawing and freezing cycles before
isolating the DNA should be avoided. If cryoprecipitates (formed during thawing of frozen
samples) are visible avoid aspirating them. Various different primary tubes, blood collection
system (e.g. Sarstedt, Greiner) and anticoagulants (except heparin) can be used to collect blood
samples for the InviMag® Blood DNA Mini Kit/ KFmL procedure.
Buffy coat is a whole-blood fraction of enriched leukocyte cells. To prepare and extract of buffy
coat the following procedure is recommended: The use of a whole blood sample
(anticoagulants: EDTA, citrate, not heparin) with a sedimented cellular fraction from staying
overnight at 4°C is recommended. The resulting bright mid-section overlaid by the clear plasma
is the buffy coat containing concentrated leukocytes that can be easily distinguished from the
erythrocytes in the bottom layer. An enrichment factor of 10 is expected from such a procedure.
Due to the enriched leukocyte content be aware to avoid overloading the purification system.
CSF (Cerebrospinal fluid) and Bone marrow:
Best results are obtained with fresh material which can be stored for 2-3 h at 4°C or for longterm storage freeze the sample at -20°C. However, often the sample is dried. Dried samples
have to be stored at 4°C in a dry surrounding.
Swabs
The protocol works with fresh prepared swabs as well as with dried swabs. Please note, that
stored and dried swabs can lead to samples that show an apoptotic DNA ladder (visible on
agarose gel as typical apoptotic DNA banding pattern).The protocol has not been validated for
isolation of DNA from swabs which are stored in special storage buffers from other providers.
STRATEC Molecular will be released of its responsibilities if other sample materials than
described in the Intended Use are processed or if the sample preparation protocols are changed
or modified.
Principle and procedure
The InviMag® Blood DNA Mini Kit/ KFmL procedure comprises following steps:
○
○
○
○
lysis of sample material and protein digestion
binding the genomic DNA to the magnetic beads
washing the beads and elimination of ethanol
elution of genomic DNA
After lysis, the DNA binds to the magnetic beads whereas contaminations and inhibitors are
efficiently removed during the following three wash steps. Highly purified DNA is eluted in
Elution Buffer.
This manual contains 4 protocols (page 13-14).
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InviMag Blood DNA Mini Kit/ KFmL 0515
Lysis
Lysis is performed at elevated temperatures in the presence of Lysis Buffer A and Proteinase
K. In case of large sample numbers we recommend the preparation of a master mix with a
volume 5% greater than that required. Carefully mix the master mix carefully prior to use!
Binding of the genomic DNA
After adding Binding Buffer B6 and SNAP Solution to the lysate the DNA is bound to the
surface of the magnetic beads.
Removing residual contaminants
Contaminants are efficiently washed away using Wash Buffer I and II, while the DNA remains
bound to the magnetic beads.
Elution
The DNA is eluted in 200 µl Elution Buffer. The eluted DNA is ready-to-use in different
subsequent downstream applications e.g. for PCR amplification, digestion with restriction
enzymes, Southern hybridizations, HLA typing etc.
Yield and quality of genomic DNA
The amount of purified DNA in the InviMag® Blood DNA Mini Kit/ KFmL procedure from whole
blood depends on the leucocytes content, sample source, transport, storage, and sample age.
Typically, a 200 μl sample of whole blood cells (samples with elevated white blood cell (WBC
counts), ranging from 3x106 to 1x107 cells/ml) from a healthy individual will yield 3–12 μg of
DNA. The typical yield usually derived from the InviMag® Blood DNA Mini Kit/ KFmL is about
3-8 µg of DNA. If the whole blood sample is mixed with anticoagulant containing buffers the
overall leukocyte concentration decreases and the yield of the DNA extraction procedure is
reduced.
Yield and quality of isolated genomic DNA is suitable for any molecular-diagnostic detection
system. The diagnostic tests should be performed according to manufacturers’ specifications.
Reagents and equipment to be supplied by user
○
○
○
○
○
○
○
Measuring cylinder (250 ml)
Pipette and pipette tips
Disposable gloves
Reaction tubes (1.5 ml / 2.0 ml)
dd H2O
Vortexer
96-100% ethanol
○ Isopropanol*
The InviMag® Blood DNA Mini Kit/ KFmL is validated with 2-Propanol; Rotipuran >99.7%, p.a.,
ACS, ISO (Order no. 6752) from Carl Roth
* Possible suppliers for Isopropanol:
Carl Roth
2-Propanol
Rotipuran >99.7%, p.a., ACS, ISO
Order no. 6752
Applichem
2-Propanol für die Molekularbiologie
Order no. A3928
10
Sigma
2-Propanol
Order no. 59304-1L-F
®
InviMag Blood DNA Mini Kit/ KFmL 0515
Important points before starting a protocol
Immediately upon receipt of the product, inspect the product and its components as well as the
package for any apparent damages, correct quantities and quality. If there are any
unconformities you have to notify STRATEC Molecular in writing with immediate effect upon
inspection thereof. If buffer bottles are damaged, contact the STRATEC Molecular Technical
Services or your local distributor. In case of liquid spillage, refer to “Safety Information” (see
page 7). Do not use damaged kit components, since their use may lead to poor kit performance.
o Always change pipet tips between liquid transfers. To avoid cross-contamination we
recommend the use of aerosol-barrier pipet tips.
o All centrifugation steps are carried out at room temperature.
o When working with chemicals, always wear a suitable lab coat, disposable gloves and
protective goggles.
o Discard contaminated gloves immediately.
o Do not combine components of different kits, unless the lot numbers are identical.
o Avoid microbial contamination of the kit reagents.
o To minimize the risk of infections from potentially infectious material, we recommend
working under laminar air-flow until the samples are lysed.
o This kit should only be used by trained personnel.
Preparing reagents and buffers
Before starting a run, equilibrate all reagents at room temperature. Where necessary, gently mix
and redissolve any precipitates by incubation at 30°C. Swirl gently to avoid foaming.
Lysis Buffer A and Elution Buffer are ready-to-use.
Add the required amount of dd-H2O (see Kit Contents, page 3) to the reaction tube containing
Proteinase K. Vortexing for 5 sec and store diluted Proteinase K at -20°C.
15 DNA-extractions:
Add 7 ml 99.7% Isopropanol to the Binding Buffer B6. Mix by intensive shaking by inverting for 1
min. Shortly before use mix by inverting several times.
Dilute Proteinase K by addition of 0.5 ml ddH2O, mix thoroughly until completely dissolving and store at
-20°C!
Add 7.5 ml of 96-100% ethanol to the bottle Wash Buffer I.
Add 42 ml of 96-100% ethanol to the bottle Wash Buffer II.
75 DNA-extractions:
Add 21 ml 99.7% Isopropanol to each Binding Buffer B6. Mix by intensive shaking by inverting for 1
min. Shortly before use mix by inverting several times.
Dilute Proteinase K by addition of 2 ml of ddH2O, mix thoroughly until completely dissolving and store
at -20°C!
Add 30 ml of 96-100% ethanol to the bottle Wash Buffer I.
Add 105 ml of 96-100% ethanol to the bottle Wash Buffer II.
Mix thoroughly and keep the bottle always firmly closed!
300 DNA-extractions:
Add 56 ml 99.7% Isopropanol to each Binding Buffer B6. Mix by intensive shaking by inverting for 1
min. Shortly before use mix by inverting several times.
Dilute Proteinase K by addition of 7 ml ddH2O, mix thoroughly until completely dissolving and store at 20°C!
Add 80 ml of 96-100% ethanol to each bottle Wash Buffer I, mix thoroughly and always keep the bottle
firmly closed!
Add 140 ml of 96-100% ethanol to each bottle Wash Buffer II, mix thoroughly and always keep the bottle
firmly closed
11
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InviMag Blood DNA Mini Kit/ KFmL 0515
Scheme of the InviMag® Blood DNA Mini Kit/ KFmL
Please carefully read the protocols before starting a run
-
If necessary add required amount of dd-H2O or 1x PBS to adjust
the sample volume to 200 µl.
Transfer 200 µl sample into a 1.5 ml reaction tube (not provided)
Add 200 µl Lysis Buffer A and 20 µl Proteinase K.
Mix by pipetting up and down or by moving up and down the
magnetic rods
Incubate the samples at 56°C for 10 min while shaking
During lysis, prefill the KingFisher Tube strip(s) with the required
buffers and appropriate volumes
Tube A:
Tube B:
Tube C:
Tube D:
Tube E:
Add 400 µl Binding Buffer B6 and 20 µl SNAP Solution
Add 800 µl Wash Buffer I.
Add 800 µl Wash Buffer II.
Add 800 µl Wash Buffer II.
Add 200 µl Elution Buffer.
Addition of 400 µl Binding Buffer B6 (follow preparing instructions)
and 20 µl SNAP Solution with the lysate
DNA binds to magnetic particles
Magnetic particle separation
3x Washing of the particle fixed DNA (3x 800 µl)
Magnetic particle separation
Elution of genomic DNA in 200 µl Elution Buffer
Magnetic Separation
Pure DNA
12
®
InviMag Blood DNA Mini Kit/ KFmL 0515
Protocol 1: Isolation of genomic DNA from up to 200 µl of whole blood
or 1–30 µl buffy coat
Please read the instructions carefully and conduct the prepared procedure.
Attention: Please be aware, that you have to prepare the Binding Buffer B6 – see instruction page: 11
Important Note: The protocol has been optimized for the isolation of genomic DNA from 200 µl of whole
blood or 30 µl buffy coat. For samples with a smaller volume than 200 µl, please fill up
to a total volume of 200 µl with distilled water or 1x PBS.
External Sample lysis in a 1.5 ml reaction tube (not provided)
1. Transfer 200 µl of whole blood or 30 µl buffy coat into a 1.5 ml reaction tube. If the
sample volume is lower than 200 µl, equilibrate with sterile water or 1x PBS.
2. Add 200 µl Lysis Buffer A and 20 µl Proteinase K.
Important Note: Vortex the sample for 10 s! An incomplete mixing will reduce the quality and yield of the
isolated DNA.
3. Incubate the sample at 56°C for 10 min while continuously shaking.
4. During lysis, prefill all tubes of the KingFisher tube strips with the required buffers and
appropriate volumes (see “Starting a Run”, page 16).
5. After lysis, transfer the sample(s) carefully into the Tube A of the corresponding
KingFisher tube strip prefilled with 400 µl Binding Buffer B6 and 20 µl SNAP Solution
Important Note:
Vortex the SNAP Solution vigorously before use!
6. Continue with “Starting a run” on page 16
Protocol 2: Isolation of genomic DNA from up to 25 µl of non
mammalian blood
Please read the instructions carefully and conduct the prepared procedure.
Attention: Please be aware, that you have to prepare the Binding Buffer B6 – see instruction page: 11
Important Note: For samples which have a smaller volume than 200 µl please adjust to a total volume of
200 µl using 1x PBS or distilled water.
If bird (e. g. chicken) or fish blood should be used that contain nucleated erythrocytes, the use of
only 10-15 µl of starting material is recommended.
I. Sample Lysis
1. Transfer max. 25 µl of non-mammalian blood (not heparin stabilized) into a 1.5 ml reaction
tube. Adjust the sample volume to 200 µl using 1x PBS or distilled water.
2. Add 200 µl Lysis Buffer A and 20 µl Proteinase K.
Important Note: Vortex the sample for 10 s! An incomplete mixing step will reduce quality and yield of the
isolated DNA.
3. Incubate the sample at 56°C for 25 min while continuously shaking.
4. During lysis, prefill all tubes of the KingFisher mL with required buffers and appropriate
volumes (see “Starting a Run”, page 16).
5. After lysis, transfer the sample carefully into Tube A of the KingFisher tube strip prefilled with
400 µl Binding Buffer B6 and 20 µl SNAP Solution (see “Starting a Run”, page 16).
13
®
InviMag Blood DNA Mini Kit/ KFmL 0515
Protocol 3: Isolation of genomic DNA from CSF and bone marrow
Please read the instructions carefully and conduct the prepared procedure.
Attention: Please be aware, that you have to prepare the Binding Buffer B6 – see instruction page: 11
Important Note: For samples which have a smaller volume than 200 µl please adjust to a total volume of
200 µl using 1x PBS or distilled water
Preparation of the starting material:
Fresh material:
○
○
1–200 µl fresh cerebrospinal fluid
1-20 µl bone marrow
Dried material (for example on hematological slides):
○
○
○
○
Moisten the dried material with a drop of PBS.
Add 180 µl PBS to a 1.5 ml microcentrifuge tube (not provided).
Scrape cytological material into the microcentrifuge tube using the edge of a clean slide.
Dissolve the resulting sludge by pipetting up and down.
I. Sample Lysis
1. Transfer the starting material into a 1.5 ml reaction tube. If the sample volume is lower than
200 µl, adjust with 1 x PBS Buffer or distilled water.
2. Add 200 µl Lysis Buffer A and 20 µl Proteinase K.
Important Note: Vortex the sample for 10 s! An incomplete mixing will reduce quality and yield of the
isolated DNA.
3. Incubate the sample at 56°C for 20 min while continuously shaking.
4. During lysis, prefill all tubes of the KingFisher mL with needed buffers and appropriate
volumes (see “Starting a Run”, page 16).
5. After the lysis transfer the sample carefully into the Tube A of the KingFisher tube strip
prefilled with 400 µl Binding Buffer B6 and 20 µl SNAP Solution (see “Starting a Run”, page
16).
Important Note: Vortex the SNAP Solution vigorously before use!
Protocol 4: Isolation of genomic DNA from swabs or rinsed liquid from
swabs
Please read the instructions carefully and conduct the prepared procedure.
Attention: Please be aware, that you have to prepare the Binding Buffer B6 – see instruction page: 11
Important Note: For samples which have a smaller volume than 200 µl, please fill up to a total volume of
200 µl with 1x PBS or distilled water. If chicken or fish blood is used a final sample
volume of 10 µl is recommended.
14
®
InviMag Blood DNA Mini Kit/ KFmL 0515
Fresh or dried swabs
I. Sample Lysis
1. Add 180 µl PBS-Buffer or distilled water to 1.5 ml reaction tube (not provided). Transfer the
swab into the tube and incubate for 3 min. Afterwards add 200 µl Lysis Buffer A and 20 µl
Proteinase K. Mix by pipetting up and down (5 times).
2. Incubate the reaction tube at 56°C for 20 min while continuously shaking on a thermomixer.
Important Note: To get maximum yield of DNA, it is essential to leave the swab inside the tube during the
complete lysis time. It is possible to cut-off the shaft of the swab to allow closing of
reaction tube cap. The removing of the swab from the reaction tube ahead of time will
lead to a dramatically reduced final yield!
3. During lysis, prefill all tubes of the KingFisher mL with required buffers and appropriate
volumes (see “Starting a Run”, page 16).
4. After lysis, carefully squeeze out the swab inside the tube wall and discard the swab.
Transfer the sample carefully into Tube A of the KingFisher tube strip prefilled with 400 µl
Binding Buffer B6 and 20 µl SNAP Solution (see “Starting a Run”, page 16).
Important Note: Vortex the SNAP Solution vigorously before use!
Swabs delivered in transportation media
Important: If the swab is delivered in stabilization media, ensure that these media are compatible with the
STRATEC Molecular chemistry. For more information contact STRATEC Molecular:
phone + 49 30 9489 2907
1. Transfer 200 µl of the transportation media into a 1.5 ml reaction tube. If the sample volume
is lower than 200 µl, equilibrate with 1x PBS or distilled water.
2. Add 200 µl Lysis Buffer A and 20 µl Proteinase K.
Important Note: Vortex the sample for 10 s! An incomplete mixing will reduce quality and yield of the
isolated DNA.
3. Incubate the sample at 56°C for 20 min while continuously shaking.
4. During lysis, prefill all tubes of the KingFisher mL with required buffers and appropriate
volumes (see “Starting a Run”, page 16).
5. After the lysis transfer the sample carefully into the Tube A of the KingFisher tube strip
prefilled with 400 µl Binding Buffer B6 and 20 µl SNAP Solution (see “Starting a Run”, page
16).
Use of the rinsed liquid from the swab
1. If the swab is delivered without transport media, rinse each swab in a 1.5 ml reaction tube
with 200–500 µl cooled disatilled water or 1x PBS. Mix for several minutes by shaking.
2. Follow the protocols for sample lysis found in the previous section “Swab delivered in
transportation media” and use 200 µl of the rinsed liquid as starting material.
15
®
InviMag Blood DNA Mini Kit/ KFmL 0515
Starting a run - Preliminary Steps to process the sample onto the
KingFisher System
Important: For working with the KingFisher mL instrument, please read carefully the manufacturer’s
documents!
1. During the sample lysis prefill the tubes of the KingFisher tube strips with the following buffers
and appropriate volumes:
Tube A:
Important:
Add 400 µl Binding Buffer B6 (follow preparing instructions) and 20 µl SNAP
Solution. After lysis, transfer the sample into tube A of the KFmL stripe
Mix the bottle with SNAP Solution carefully by vigorously shaking or vortexing !
Tube B:
Add 800 µl Wash Buffer I.
Tube C:
Add 800 µl Wash Buffer II.
Tube D:
Add 800 µl Wash Buffer II.
Tube E:
Add 200 µl Elution Buffer.
2. Place the prefilled KingFisher tube strips with the tube tray into the KingFisher system
3. Place the KingFisher tip combs into the slots of the instrument!
4. Choose KFmL assay file “InviMag Blood DNA KFmL” and press the “START” button
The following extraction steps run automatically on the KFmL system:
Binding of the DNA
Automatically sample mixing for 5 minutes. MAP separation. Moving of the MAP into well B.
First Washing
Automatically sample mixing for 90 s. MAP separation. Moving of the MAP into well C.
Second Washing
Automatically sample mixing for 1 min. MAP separation. Moving of the MAP into well D.
Third Washing and Drying
Automatically sample mixing for 1 min. MAP separation. Drying the MAP outsight the Tube for
5 min. Moving of the MAP into the well E.
Elution of the DNA
Incubation of the MAP into the Tube E for 15 minutes by mixing. MAP separation. The MAP will
then be automatically removed into well D (disposal).
Important Note:
After finishing the extraction protocol, the Tube E contains the extracted genomic DNA
Store the DNA under adequate conditions. We recommend to transfer the extracted DNA
into 1.5 ml reaction tubes for further storage and freeze the DNA at –20°C.
If the extracted DNA contains carryover of magnetic particles, transfer the DNA into a
1.5 ml reaction tube, centrifuge at maximum speed for 1 min and transfer the DNAcontaining supernatant into a new tube.
16
®
InviMag Blood DNA Mini Kit/ KFmL 0515
For self programming the KFmL system
[ PROTOCOL PROPERTIES ]
Name = InviMag Blood DNA Mini Kit/ KFmL
Protocol template version = 3.1
Instrument type = KFmL
Description = KFmL protocol for isolation of
genomic DNA from blood samples with the
InviMag Blood DNA Mini Kit/ KFmL
Washing_1
Plate: Blood Mini (B)
Beginning of step:
Precollect: No
Release time [hh:mm:ss]: 00:00:10
Release speed: Fast
[ PLATE LAYOUTS ]
Mixing/pause parameters:
Pause for manual handling: No
Mixing time [hh:mm:ss]: 00:01:30
Mixing speed: Fast
Binding plate
Plate type = KingFisher tub strip 1000µl
Plate change message = Insert Bind plate
- volume = 200, name = Sample
- volume = 200, name = Lysis Buffer A
- volume = 20, name = Proteinase K
End of step:
Postmix: No
Collect count: 3
Collect time [s]: 2
Washing plate_1
Plate type = KingFisher tub strip 1000µl
Plate change message = Insert Wash 1
- volume = 800, name = Wash buffer I
Washing_2
Plate: Blood Mini (C)
Beginning of step:
Precollect: No
Release time [hh:mm:ss]: 00:00:10
Release speed: Fast
Washing plate_2
Plate type = KingFisher tub strip 1000µl
Plate change message = Insert Wash 2
- volume = 800, name = Wash buffer II
Washing plate_3
Plate type = KingFisher tub strip 1000µl
Plate change message = Insert Wash 3
- volume = 800, name = Wash buffer II
Mixing/pause parameters:
Pause for manual handling: No
Mixing time [hh:mm:ss]: 00:01:00
Mixing speed: Fast
Elution plate
Plate type = KingFisher tub strip 1000µl
Plate change message = Insert Elution
- volume = 200, name = Elution Buffer
End of step:
Postmix: No
Collect count: 3
Collect time [s]: 2
[ STEPS ]
Binding
Plate: Blood Mini (A)
Washing_3
Plate: Blood Mini (D)
Beginning of step:
Precollect: No
Release time [hh:mm:ss]: 00:00:10
Release speed: Fast
Beginning of step:
Precollect: No
Release time [hh:mm:ss]: 00:00:10
Release speed: Fast
Mixing/pause parameters:
Pause for manual handling: No
Mixing time [hh:mm:ss]: 00:01:00
Mixing speed: Fast
End of step:
Postmix: No
Collect count: 3
Collect time [s]: 2
Mixing/pause parameters:
Pause for manual handling: No
Mixing time [hh:mm:ss]: 00:05:00
Mixing speed: Slow
End of step:
Postmix: No
Collect count: 4
Collect time [s]: 3
Drying
17
®
InviMag Blood DNA Mini Kit/ KFmL 0515
Mixing time [hh:mm:ss]: 00:10:00
Mixing speed: Slow
Plate: Washing Plate 4
Dry time [hh:mm:ss]: 00:05:00*
Tip position: Outside well / tube
End of step:
Postmix: No
Collect count: 4
Collect time [s]: 3
Elution
Plate : Blood Mini (E)
Beginning of step:
Precollect: No
Release time [hh:mm:ss]: 00:00:10
Release speed: Medium
Remove_Beads
Plate: Blood Mini (D)
Release time [hh:mm:ss]: 00:00:30
Release speed: Fast
Mixing/pause parameters:
Pause for manual handling: No
18
®
InviMag Blood DNA Mini Kit/ KFmL 0515
Troubleshooting
Problem
Probable cause
Comments and suggestions
low amount of extracted
DNA
insufficient lysis
increase lyses time
Reduce amount of starting material
incomplete elution
take higher volume of Elution
Buffer, be sure you pipet the
Elution Buffer with the right amount
to the right position
low amount of SNAP Solution
mix SNAP Solution thoroughly
before pipetting to the KingFisher
tube
too much Elution Buffer
elute the DNA with lower volume of
Elution Buffer
incorrect storage of starting
material
ensure that the storage of starting
material was correctly
Avoid thawing of the material
incorrect storage of starting
material
ensure that the storage of starting
material was correctly
Avoid thawing of the material
old material
ensure that the starting material is
fresh or stored under appropriate
condition (for long time storage at –
20°C)
avoid thawing and freezing of the
material
old material often contains
degraded DNA
DNA does not perform
well in downstreamapplications (e.g. realtime PCR or PCR)
ethanol carryover during elution
increase drying time for removing
of ethanol
salt carryover during elution
check up the Wash Buffers for salt
precipitates, if there are any
precipitates, solve these
precipitates by careful warming
ensure that the Wash Buffers are
at room temperature
low A260:A280 ratio from
UV measurement, eluted
DNA is brown colored
small part of the magnetic particles
are left in the elution
centrifuge down at full speed for 1
min and transfer supernatant to a
new tube
low concentration of
extracted DNA
degraded or sheared
DNA
19
®
InviMag Blood DNA Mini Kit/ KFmL 0515
Appendix
KingFisher Software 3.1
The KingFisher Software 3.1 was used to create assay files for the KFmL, KF96 and KFflex96
instruments. The respective assay file can either be transferred onto the KingFisher workstation
or be started directly from within the BindIt software. Keep in mind that directly run assay files
are not stored in the workstation memory!
Note: When creating assay files for usage with KingFisher instruments in combination with Microtiter
Deep Well plates (e.g. Thermo Electron), it is essential to use the KingFisher software 3.1 for assay
development as this software version includes the correct adjustments for this plate. It is highly
recommended to use Thermo Microtiter Deep Well plates with KF96 / KFflex96 workstations to ensure the
best purification result.
PC requirements for KingFisher Software 3.1
PC requirements
Interface
Serial communication port via a RS-232 full duplex interface
Supported operating systems
Microsoft Windows 2000
Microsoft Windows XP Professional
Disk space
500 MB free disk space
Processor
Intel Pentium ≥ 700 MHz recommended
Memory
220 MB RAM recommended
Serial ports available
1
Pointing device
Mouse or equivalent is necessary
CD-ROM drive
1
Monitor / color settings
SVGA monitor with at least 1024 x 768 resolution and at least
a 16-bit color environment
Service packs installed
Microsoft Windows 2000: Service Pack 4 (or greater)
Microsoft Windows XP Professional: Service Pack 2 (or
greater)
Browser
Microsoft Internet Explorer 6.0 (or greater) installed
If you do not have the correct Service Packs installed, you can download them from the
Microsoft web pages: http://www.microsoft.com/
20
®
InviMag Blood DNA Mini Kit/ KFmL 0515
General notes on handling DNA
Starting material
This kit is designed for extraction of DNA from blood, but even human blood is different between
individuals depending on age, health, and conditions of life. If you are using blood from animals
keep in mind that lyses conditions of blood differ depending on the species. Also remember that
non-mammalian blood contains erythrocytes with nuclei. So for special applications adaptation
of starting volumes and lyses time may be recommended.
Nature of DNA
The length and delicate physical nature of DNA requires careful handling to avoid damage due
to shearing and enzymatic degradation. Other conditions that affect the integrity and stability of
DNA include acidic and alkaline environments, high temperature, and UV irradiation. Careful
isolation and handling of high molecular weight DNA is necessary to ensure compatibility with
various downstream applications. Damaged DNA could perform poorly in applications such as
genomic Southern blotting, long-template PCR.
Storage of DNA
A working stock of DNA can be stored at 2–4˚C for several weeks. For long-term storage DNA
should be stored at -20˚C, but storing at -20°C can cause shearing, particularly if the DNA is
exposed to repeated freeze-thawing cycles.
Note that the solution in which the nucleic acid is eluted in, will affect the stability during storage.
Pure water lacks buffering capacity and an acidic pH may lead to acid hydrolysis. Tris or TrisEDTA buffer contains sufficient buffering capacity to prevent acid hydrolysis.
Drying, dissolving and pipetting DNA
Avoid over drying genomic DNA after ethanol precipitation. It is better to let it air dry than to use
a vacuum, although vacuum drying can be used with caution.
Avoid vigorous pipetting. Pipetting genomic DNA through small tip openings causes shearing or
nicking. One way to decrease shearing of genomic DNA is to use special tips that have wide
openings designed for pipetting genomic DNA.
DNA Yield
The amount of purified DNA from the whole blood depends on the leucocytes content, sample
source, transport, storage, and age. Various different primary tubes and anticoagulants (except
heparin) can be used to collect blood samples for the Invisorb® procedure.
21
®
InviMag Blood DNA Mini Kit/ KFmL 0515
Ordering information
Product
InviMag® Blood DNA Mini Kit/ KFmL
InviMag® Blood DNA Mini Kit/ KFmL
InviMag® Blood DNA Mini Kit/ KFmL
InviMag® Blood DNA Mini Kit/ KF96
InviMag® Blood DNA Mini Kit/ KF96
Package Size
Catalogue No.
15 preparations
75 preparations
300 preparations
2431110100
2431110200
2431110400
1 x 96 preparations
5 x 96 preparations
7431300100
7431300200
Invisorb® Spin Blood Mini Kit
Invisorb® Spin Blood Mini Kit
50 preparations
250 preparations
1031100200
1031100300
Invisorb® Spin Blood Midi Kit
50 preparations
1031110200
Invisorb® Blood Universal Kit
Invisorb® Blood Universal Kit
50 ml
500 ml
Invisorb® Blood Mini HTS 96 Kit/ C
Invisorb® Blood Mini HTS 96 Kit/ C
1031150100
1031150200
4 x 96 preparations
24 x 96preparations
7031300300
7031300400
4 x 96 preparations
24 x 96 preparations
7131310300
7131410400
using a centrifuge
Invisorb® Blood Mini HTS 96 Kit/ X
Invisorb® Blood Mini HTS 96 Kit/ X
using the X-tractor Gene™, Corbett Robotics
Single Components for InviMag® Blood DNA Mini Kit
Lysis Buffer A
SNAP Solution
Binding Buffer B6
Elution Buffer
Wash Buffer I
Wash Buffer II
30 ml
1.5 ml
30 ml
30 ml
30 ml
60 ml
7431301100
7431305200
7431302100
7431304000
7431303300
7431303400
Ordering information (KingFisher mL and consumables from Thermo Scientific)
Cat.no
5400050
97002111
97002121
97002131
97002141
Description
KingFisher mL, Magnetic Particle Processor, 100-240 V, 50/60 Hz
KingFisher mL tip comb, 800 pcs
KingFisher mL tube, 900 pcs (20x45 pcs)
KingFisher mL Combi 60 (tubes and tip combs for 60 samples)
KingFisher mL Combi 240 (tubes and tip combs for 240 samples)
Possible suppliers for Isopropanol:
Carl Roth
2-Propanol
Rotipuran >99.7%, p.a., ACS, ISO
Order no. 6752
Applichem
2-Propanol für die Molekularbiologie
Order no. A3928
22
Sigma
2-Propanol
Order no. 59304-1L-F
®
InviMag Blood DNA Mini Kit/ KFmL 0515
STRATEC Molecular GmbH
Robert-Rössle-Str. 10
13125 Berlin, Germany
www.stratec.com
1G3a01/05/2015
Phone: +49 30 94 89 29 01
Fax: +49 30 94 89 29 09
E-mail: [email protected]